Metabolic profile of blood serum in experimental arterial hypertension.
The etiology of essential hypertension is intricate, since it employs simultaneously various body systems related to the regulation of blood pressure in one way or another: the sympathetic nervous system, renin-angiotensin-aldosterone and hypothalamic-pituitary-adrenal systems, renal and endothelial mechanisms. The pathogenesis of hypertension is influenced by a variety of both genetic and environmental factors, which determines the heterogeneity of the disease in human population. Hence, there is a need to perform research on experimental models - inbred animal strains, one of them being ISIAH rat strain, which is designed to simulate inherited stress-induced arterial hypertension as close as possible to primary (or essential) hypertension in humans. To determine specific markers of diseases, various omics technologies are applied, including metabolomics, which makes it possible to evaluate the content of low-molecular compounds - amino acids, lipids, carbohydrates, nucleic acids fragments - in biological samples available for clinical analysis (blood and urine). We analyzed the metabolic profile of the blood serum of male ISIAH rats with a genetic stress-dependent form of arterial hypertension in comparison with the normotensive WAG rats. Using the method of nuclear magnetic resonance spectroscopy (NMR spectroscopy), 56 metabolites in blood serum samples were identified, 18 of which were shown to have significant interstrain differences in serum concentrations. Statistical analysis of the data obtained showed that the hypertensive status of ISIAH rats is characterized by increased concentrations of leucine, isoleucine, valine, myo-inositol, isobutyrate, glutamate, glutamine, ornithine and creatine phosphate, and reduced concentrations of 2-hydroxyisobutyrate, betaine, tyrosine and tryptophan. Such a ratio of the metabolite concentrations is associated with changes in the regulation of glucose metabolism (metabolic markers - leucine, isoleucine, valine, myo-inositol), of nitric oxide synthesis (ornithine) and catecholamine pathway (tyrosine), and with inflammatory processes (metabolic markers - betaine, tryptophan), all of these changes being typical for hypertensive status. Thus, metabolic profiling of the stress-dependent form of arterial hypertension seems to be an important result for a personalized approach to the prevention and treatment of hypertensive disease.


The epidemic of AIDS: a failure of public health policy.
The AIDS epidemic was engrafted on a substantial and rising epidemic of sexually transmitted diseases. Complex social, political, and professional factors are examined through a single risk group to illuminate the initial and continuing failure to combat the epidemic. Selective traditional public health control measures, applied vigorously and creatively within a new legal framework, are still appropriate.


A Human Digital-Twin-Based Framework Driving Human Centricity towards Industry 5.0.
This work presents a digital-twin-based framework focused on orchestrating human-centered processes toward Industry 5.0. By including workers and their digital replicas in the loop of the digital twin, the proposed framework extends the traditional model of the factory's digital twin, which instead does not adequately consider the human component. The overall goal of the authors is to provide a reference architecture to manufacturing companies for a digital-twin-based platform that promotes harmonization and orchestration between humans and (physical and virtual) machines through the monitoring, simulation, and optimization of their interactions. In addition, the platform enhances the interactions of the stakeholders with the digital twin, considering that the latter cannot always be fully autonomous, and it can require human intervention. The paper also presents an implemented scenario adhering to the proposed framework's specifications, which is also validated with a real case study set in a factory plant that produces wooden furniture, thus demonstrating the validity of the overall proposed approach.


Intergenerational and Social Interventions to Improve Children's Oral Health.
Dental caries and gingival and periodontal diseases are commonly occurring, preventable chronic conditions in children. These diseases are more common in disadvantaged communities and marginalized populations. Thus, public health approaches that stress prevention are key to improving oral health equity. There is currently limited evidence on which community-based, population-level interventions are most effective and equitable in promoting children's oral health. More rigorous measurement and reporting of study findings are needed to improve the quality of available evidence. Improved understanding of the multilevel influences of children's oral health may lead to the design of more effective and equitable social interventions.


Distance from Primary Tumor Is the Strongest Predictor for Early Onset of Brain Metastases in Melanoma.
The frequency of brain metastasis (BM) is up to 45-50% in patients with advanced melanoma. Our aim was to identify the risk factors for the early occurrence of BM. A total of 333 patients with BM were identified from our database of 2,972 patients with melanoma between 2003-2015. The median elapsed time to BM (TTBM) was significantly associated with Breslow thickness, ulceration, location, and patient age. Head and neck location was the strongest predictor for early BM development [hazard ratio (HR)=1.81, 95% confidence interval (CI)=1.05-3.12; p=0.031) followed by Breslow thickness >2 mm (HR=1.53, 95% CI=1.04-2.23; p=0.027). Body part-specific median TTBM was 51.5, 43, 38.5, 32, 35, 36.5, 35.5 and 19 months in leg-foot, thigh, abdomen-pelvic, chest-back, lower arm-hand, upper arm-shoulder, face-neck and scalp regions, respectively. We suggest brain magnetic resonance imaging follow-up in the high-risk patient group of patients with melanoma in the head and neck region, especially for those with primary melanoma over Breslow 2 mm located in the scalp.


Comparison of Antemortem and Environmental Samples for Zebrafish Health Monitoring and Quarantine.
Molecular diagnostic assays offer both exquisite sensitivity and the ability to test a wide variety of sample types. Various types of environmental sample, such as detritus and concentrated water, might provide a useful adjunct to sentinels in routine zebrafish health monitoring. Similarly, antemortem sampling would be advantageous for expediting zebrafish quarantine, without euthanasia of valuable fish. We evaluated the detection of Mycobacterium chelonae, M. fortuitum, M. peregrinum, Pseudocapillaria tomentosa, and Pseudoloma neurophilia in zebrafish, detritus, pooled feces, and filter membranes after filtration of 1000-, 500-, and 150-mL water samples by real-time PCR analysis. Sensitivity varied according to sample type and pathogen, and environmental sampling was significantly more sensitive than zebrafish sampling for detecting Mycobacterium spp. but not for Pseudocapillaria neurophilia or Pseudoloma tomentosa. The results of these experiments provide strong evidence of the utility of multiple sample types for detecting pathogens according to each pathogen's life cycle and ecological niche within zebrafish systems. In a separate experiment, zebrafish subclinically infected with M. chelonae, M. marinum, Pleistophora hyphessobryconis, Pseudocapillaria tomentosa, or Pseudoloma neurophilia were pair-spawned and individually tested with subsets of embryos from each clutch that received no rinse, a fluidizing rinse, or were surface-disinfected with sodium hypochlorite. Frequently, one or both parents were subclinically infected with pathogen(s) that were not detected in any embryo subset. Therefore, negative results from embryo samples may not reflect the health status of the parent zebrafish.


Non-enhanced CT Maximum Intensity Projections for the Detection of Large Vessel Occlusions.
Identification of large vessel occlusions (LVO) is important with recent guidelines supporting endovascular therapy in selected acute ischemic stroke patients. Many stroke centers perform CT angiography (CTA) in patients with suspected LVO, however this requires additional time and contrast administration. Non-enhanced CT maximum intensity projection (NECT-MIPs) may offer a rapid alternative to CTA. We retrospectively reviewed acute stroke patients with LVO in the UCSD Stroke Registry, presenting between 6/2014-7/2016. NECT-MIPs were evaluated for presence of LVO. Gold standard comparison was to CTA. Results were stratified by level of training (Faculty, Fellow and Acute Care Practitioners [ACPs]). Inter-rater agreement was assessed using Fleiss' Kappa Coefficient. We reviewed 24 patients using NECT-MIPs for the detection of LVO. Faculty had a sensitivity and specificity of 95% & 92% for ICA/M1, 42% & 100% for M2, and 67% & 96% for basilar occlusions. Fellows and ACPs had a sensitivity and specificity of 61% & 94% for ICA/M1, 19% & 83% for M2, and 75% & 95% for basilar occlusions. Inter-rater agreement among Faculty readers was k=0.75 for ICA/M1, k=0.79 for M2 and k=0.14 for basilar occlusions. Among Fellows and ACPs, k=0.57 for ICA/M1, k=0.40 for M2, and k=0.27 for basilar occlusions. NECT-MIPs have high sensitivity and specificity for the detection of LVO when compared to CTA. Inter-rater agreement is fair and higher amongst more experienced reviewers. These results suggest that NECT-MIPs may be helpful to streamline the identification of LVO and reduce door to needle and door to intervention times.


Identification of biomarkers associated with diagnosis of postmenopausal osteoporosis patients based on bioinformatics and machine learning.
Background: Accumulating evidence suggests that postmenopausal osteoporosis (PMOP) is a common chronic systemic metabolic bone disease, but its specific molecular pathogenesis remains unclear. This study aimed to identify novel genetic diagnostic markers for PMOP. Methods: In this paper, we combined three GEO datasets to identify differentially expressed genes (DEGs) and performed functional enrichment analysis of PMOP-related differential genes. Key genes were analyzed using two machine learning algorithms, namely, LASSO and the Gaussian mixture model, and candidate biomarkers were found after taking the intersection. After further ceRNA network construction, methylation analysis, and immune infiltration analysis, ACACB and WWP1 were finally selected as diagnostic markers. Twenty-four clinical samples were collected, and the expression levels of biomarkers in PMOP were detected by qPCR. Results: We identified 34 differential genes in PMOP. DEG enrichment was mainly related to amino acid synthesis, inflammatory response, and apoptosis. The ceRNA network construction found that XIST-hsa-miR-15a-5p/hsa-miR-15b-5p/hsa-miR-497-5p and hsa-miR-195-5p-WWP1/ACACB may be RNA regulatory pathways regulating PMOP disease progression. ACACB and WWP1 were identified as diagnostic genes for PMOP, and validated in datasets and clinical sample experiments. In addition, these two genes were also significantly associated with immune cells, such as T, B, and NK cells. Conclusion: Overall, we identified two vital diagnostic genes responsible for PMOP. The results may help provide potential immunotherapeutic targets for PMOP.


Clinical characteristics of IgG4-related disease with lung involvement in 14 patients.
Immunoglobulin G4-related diseases (IgG4-RD) is a rare autoimmune disease, and there is no specific diagnostic test for patients with lung involvement yet. This study aims to summarize the clinical characteristics of IgG4-RD with lung involvement and improve the understanding and diagnosis of this disease. All patients diagnosed with IgG4-RD in the Second Xiangya Hospital from December 2014 to February 2022 were re-diagnosed according to the recommendations of Chinese Expert Consensus on the Diagnosis and Treatment of IGG4-Related Diseases in 2021. The clinical data of 14 IgG4-RD patients with pulmonary abnormalities were collected and analyzed. Among the 14 patients, 11 were males and 3 were females, and the median age of diagnosis was 66 (22-82) years old. Six cases had respiratory symptoms such as cough, sputum and short breath. Extrapulmonary involvement was the most common in the glands of head and neck (6/14), followed by pancreas and bile duct (4/14). Elevated serum IgG4 level was found in all patients, and most (11/14) were accompanied by abnormal inflammatory markers. Patients' pulmonary imaging findings were diverse, the most common performances were mediastinal/hilar lymphadenopathy (12/14), followed by multiple pulmonary nodules (9/14), patchy density enhancement (7/14) and the increased broncho vascular bundles (6/14). Lung biopsy was performed in 9 patients, their pathology results showed lymphoplasmic cell infiltration, 5 cases of them had interstitial fibrosis, 2 cases with phlebitis, and extrapulmonary biopsy was performed in 8 patients. Immunohistochemical results of all the patients showed that the number of IgG4+ plasma cells was more than 10 per high magnification, and the ratio of IgG4/IgG was more than 40%. For treatment, 12 patients received hormone therapy, and 5 patients combined immunosuppressive therapy with hormone. 10 patients were in remission after treatment, while 2 patients were progressed. IgG4-RD with lung involvement is rare and has no specific clinical manifestation. Its pulmonary imaging is diverse. Diagnosis for it should combine with serum IgG4 level and pathological examination. Glucocorticoid is the first line treatment, and combination with immunosuppressant can help prevent disease recurrence.


Human rights and maternal-fetal HIV transmission prevention trials in Africa.
The human rights issues raised by the conduct of maternal-fetal human immunodeficiency virus transmission trials in Africa are not unique to either acquired immunodeficiency syndrome or Africa, but public discussion of these trials presents an opportunity for the United States and other wealthy nations to take the rights and welfare of impoverished populations seriously. The central issue at stake when developed countries perform research on subjects in developing countries is exploitation. The only way to prevent exploitation of a research population is to insist not only that informed consent be obtained but also that, should an intervention be proven beneficial, the intervention will be delivered to the impoverished population. Human rights are universal and cannot be compromised solely on the basis of beliefs or practices of any one country or group. The challenge to the developed countries is to implement programs to improve the health of the people in developing countries both by improving public health infrastructure and by delivering effective drugs and vaccines to the people.


Effective data filtering is prerequisite for robust microbial association network construction.
Microorganisms do not exist as individual population in the environment. Rather, they form complex assemblages that perform essential ecosystem functions and maintain ecosystem stability. Besides the diversity and composition of microbial communities, deciphering their potential interactions in the form of association networks has attracted many microbiologists and ecologists. Much effort has been made toward the methodological development for constructing microbial association networks. However, microbial profiles suffer dramatically from zero values, which hamper accurate association network construction. In this study, we investigated the effects of zero-value issues associated with microbial association network construction. Using the TARA Oceans microbial profile as an example, different zero-value-treatment approaches were comparatively investigated using different correlation methods. The results suggested dramatic variations of correlation coefficient values for differently treated microbial profiles. Most specifically, correlation coefficients among less frequent microbial taxa were more affected, whichever method was used. Negative correlation coefficients were more problematic and sensitive to network construction, as many of them were inferred from low-overlapped microbial taxa. Consequently, microbial association networks were greatly differed. Among various approaches, we recommend sequential calculation of correlation coefficients for microbial taxa pairs by excluding paired zero values. Filling missing values with pseudo-values is not recommended. As microbial association network analyses have become a widely used technique in the field of microbial ecology and environmental science, we urge cautions be made to critically consider the zero-value issues in microbial data.


Use of high inoculum for early metabolic signalling and rapid susceptibility testing of Aspergillus species.
To develop and evaluate a new method for rapid susceptibility testing of Aspergillus spp. based on early metabolic signalling of high-inoculum biomass. Susceptibility to amphotericin B and voriconazole was studied in 39 clinical isolates of Aspergillus spp. (16 Aspergillus fumigatus, 11 Aspergillus flavus, 12 Aspergillus terreus). At 6 or 8 h after inoculation for A. fumigatus and A. flavus, and at 8 or 12 h after inoculation for A. terreus, 100 microg/mL of the tetrazolium salt XTT and 25 microM menadione were added and absorbance measured at 450 nm after 2 h of incubation at 37 degrees C. Inocula used were 10(6) conidia/mL for A. fumigatus and A. terreus and 10(5) conidia/mL for A. flavus, as lower inocula exhibited very low metabolic activity at these time points. Data were analysed with the sigmoid E(max) model and compared with visual (lowest drug concentration showing no growth) and spectrophotometric MIC determination at 48 h (CLSI M38-A method). The E(max) model described well the concentration-effect relationship for early metabolic activity and 48 h fungal biomass (median r(2): 0.97 and 0.93, respectively). Use of the model allowed characterization and quantification of species- and drug-related differences in pharmacological inhibition of early metabolic activity as well as calculation of appropriate cutoff levels for MIC determination with the XTT assay. Using these cutoff levels, for A. fumigatus and A. flavus at both time points (6 and 8 h) and for A. terreus at 12 h, the agreement (+/- one dilution) of the XTT assay with the CLSI method was 91-100% and its reproducibility was 97-100%. This newly developed high-inoculum-based method provides rapid and reproducible MIC determinations for Aspergillus spp.


Structural Distortion, Spin-Phonon Coupling, Interband Electronic Transition, and Enhanced Magnetization in Rare-Earth-Substituted Bismuth Ferrite.
Rare-earth ions (RE = La3+, Nd3+, and Er3+) substituted BiFeO3 (BFO) ceramics were synthesized by a conventional solid-state sintering procedure. X-ray diffraction patterns and Raman spectra confirm a rhombohedral R3c symmetry in all samples with significant distortion in FeO6 octahedron, as well as the occurrence of remarkable spin-phonon coupling and notable change in magnetic transition temperature induced by RE dopants. The enhanced magnetization was observed in all RE-doped BFO ceramics, unveiling that the spatial spin structure of BFO should be perturbed by RE dopants. Diffuse reflectance spectra show a conspicuous evolution of interband electronic transitions in RE-doped BFO ceramics. Especially, the two crystal-field d-d band transitions (6A1g→4T1 and 6A1g→4T2g) exhibit a linear red-shift behavior with the reduction in the cell volume, which is well-linked with a linear tendency of increased magnetization. On the basis of these investigations, a possible mechanism was proposed in this paper to demonstrate the correlation between the structural distortion, interband electronic transitions, and magnetic properties in RE-doped BFO ceramics.


Right Ventricular Function Improves Early After Percutaneous Mitral Valve Repair in Patients Suffering From Severe Mitral Regurgitation.
Percutaneous mitral valve edge-to-edge procedure (PMVR) using the MitraClip® system (Abbot Vascular, CA) is an established therapy for severe mitral regurgitation (MR) in patients judged inoperable or at high surgical risk. Besides determining exercise capacity, right ventricular (RV) function has prognostic value in heart failure and after cardiac surgery. We therefore investigated the impact of PMVR on RV function in patients with severe MR. Sixty-three patients undergoing PMVR at our department were prospectively enrolled. Transthoracic echocardiography was performed before, early (2-12d) after PMVR and after 3 months, including advanced echocardiographic analyses such as 3D imaging and strain analyses. At baseline, all patients presented with advanced heart failure symptoms. Etiology of MR was more often secondary and, if present, left ventricular (LV) dysfunction was predominantly caused by ischemic cardiomyopathy. PMVR substantially reduced MR to a grade ≤ 2 in most patients. Echocardiographic assessment revealed a largely unchanged LV systolic function early after PMVR, while in contrast RV function substantially improved after PMVR [3D RV EF (%): pre 33.7% [27.4; 39.6], post 40.0% [34.5; 46.0] (p < 0.01 vs. pre), 3 months 42.8% [38.3; 48.1] (p < 0.01 vs. pre); 2D RV GLS (%): pre -12.9% [-14.5; -10.5], post -16.0% [-17.9; -12.6] (p < 0.01 vs. pre), 3 months -17.2% [-21.7; -14.9] (p < 0.01 vs. pre)]. Factors that attenuated RV improvement were larger ventricular volumes, lower LV function, secondary MR, and a higher STS score (all p < 0.05). By using advanced echocardiographic parameters, we discovered an early improvement of RV function after PMVR that is preserved for months, independent from changes in LV function. Improvement of RV function was less pronounced in patients presenting with an advanced stage of heart failure and a higher burden of comorbidities reflected by the STS score.


The time course of hydroxyl radical formation following spinal cord injury: the possible role of the iron-catalyzed Haber-Weiss reaction.
This study explores whether the hydroxyl radical (*OH)-one of the most destructive reactive oxygen species-plays a role in secondary spinal cord injury (SCI). First, we measured the time course of *OH formation in rat spinal tissue after impact SCI by administering salicylate as a trapping agent into the intrathecal space of the cord and measuring the hydroxylation products of salicylate, 2,3- and 2,5-dihydroxybenzoic acid (2,3- and 2,5-DHBA) by HPLC. The 2,3-DHBA concentration was significantly higher in injured spinal tissue than in sham controls at 5 min, 1 and 3 h, but not at 5 h post-injury. Second, we generated *OH by administering H(2)O(2) and FeCl(2)/EDTA (Fenton's reagents) at the concentrations produced by SCI into the gray matter of the cord for 4 h and found that it induced significant cell loss at 24 h post-*OH exposure. Mn (III) tetrakis (4-benzoic acid) porphyrin(MnTBAP)-a broad spectrum reactive species scavenger-significantly reduced *OH-induced cell death. Finally, we generated superoxide and administered FeCl(3)/EDTA in the intrathecal space of the cord at the concentration produced by SCI and measured extracellular *OH formation in the gray matter of the cord by microdialysis sampling. We found that the levels of *OH significantly increased compared to the pre-administration level, indicating that *OH can be produced in vivo by the iron-catalyzed Haber-Weiss reaction. All together, we demonstrated that *OH is an endogenous secondary damaging agent following SCI and the metal-catalyzed Haber-Weiss reaction may contribute to early *OH formation after SCI.


Effect of hindlimb suspension and clenbuterol treatment on polyamine levels in skeletal muscle.
Polyamines are unbiquitous, naturally occurring small aliphatic, polycationic, endogenous compounds. They are involved in many cellular processes and may serve as secondary or tertiary messengers to hormonal regulation. The relationship of polyamines and skeletal muscle mass of adductor longus, extensor digitorum longus, and gastrocnemius under unloading (hindlimb suspension) conditions was investigated. Unloading significantly affected skeletal muscle polyamine levels in a fiber-type-specific fashion. Under loading conditions, clenbuterol treatment increased all polyamine levels, whereas under unloading conditions, only the spermidine levels were consistently increased. Unloading attenuated the anabolic effects of clenbuterol in predominately slow-twitch muscles (adductor longus), but had little impact on clenbuterol's action as a countermeasure in fast- twitch muscles such as the extensor digitorum longus. Spermidine appeared to be the primary polyamine involved in skeletal muscle atrophy/hypertrophy.


Fusion of building information and range imaging for autonomous location estimation in indoor environments.
We present a novel approach for autonomous location estimation and navigation in indoor environments using range images and prior scene knowledge from a GIS database (CityGML). What makes this task challenging is the arbitrary relative spatial relation between GIS and Time-of-Flight (ToF) range camera further complicated by a markerless configuration. We propose to estimate the camera's pose solely based on matching of GIS objects and their detected location in image sequences. We develop a coarse-to-fine matching strategy that is able to match point clouds without any initial parameters. Experiments with a state-of-the-art ToF point cloud show that our proposed method delivers an absolute camera position with decimeter accuracy, which is sufficient for many real-world applications (e.g., collision avoidance).


Cryogel biomaterials for neuroscience applications.
Biomaterials in the form of 3D polymeric scaffolds have been used to create structurally and functionally biomimetic constructs of nervous system tissue. Such constructs can be used to model defects and disease or can be used to supplement neuronal tissue regeneration and repair. One such group of biomaterial scaffolds are hydrogels, which have been widely investigated for cell/tissue culture and as cell or molecule delivery systems in the field of neurosciences. However, a subset of hydrogels called cryogels, have shown to possess several distinct structural advantages over conventional hydrogel networks. Their macroporous structure, created via the time and resource efficient fabrication process (cryogelation) not only allows mass fluid transport throughout the structure, but also creates a high surface area to volume ratio for cell growth or drug loading. In addition, the macroporous structure of cryogels is ideal for applications in the central nervous system as they are very soft and spongey, yet also robust, which makes them a user-friendly and reproducible tool to address neuroscience challenges. In this review, we aim to provide the neuroscience community, who may not be familiar with the fundamental concepts of cryogels, an accessible summary of the basic information that pertain to their use in the brain and nervous tissue. We hope that this review shall initiate creative ways that cryogels could be further adapted and employed to tackle unsolved neuroscience challenges.


[Collaborative innovation boosting the safe and rational use of traditional Chinese medicines].
As entering a new era,our country has introduced a series of favorable policies,which may provide the powerful and new momentum for the development of traditional Chinese medicine( TCM). However,more and more attentions have been paid to the safety and effectiveness of TCM. Tripterygium wilfordii Hook.f. is one of Chinese herbs with clinical efficacy and safety risk. In recent years,accumulating groups have carried out a series of exploratory studies on the clinical rational use of T. wilfordii-related preparations. Considering this situation,the goal of this special issue is to bring together a collection of original research and review articles addressing the expanding field of T. wilfordii. The special issue covers the clinical application,pharmacodynamics,toxicology,pharmacodynamics,resource identification and molecular pharmacognosy of T. wilfordii-related preparations. It focuses on the multi-disciplinary collaborative innovation,and may provide a stimulating resource for the fascinating subject of the safe and rational use of TCM,as well as have important practical significance and promotion value for the healthy development of TCM industry.


Adhesion of tumour-infiltrating lymphocytes to endothelium: a phenotypic and functional analysis.
Efficacy of cancer immunotherapy with cultured tumour-infiltrating lymphocytes (TILs) depends upon infused TILs migrating into tumour-bearing tissue, in which they mediate an anti-tumour response. For TILs to enter a tumour, they must first bind to tumour endothelium, and this process depends on TILs expressing and regulating the function of relevant cell-surface receptors. We analysed the cell-surface phenotype and endothelial binding of TILs cultured from human melanoma and compared them with peripheral blood T cells and with allostimulated T cells cultured under similar conditions. Compared with peripheral blood T cells, TILs expressed high levels of five integrins, two other adhesion molecules, including the skin homing molecule CLA, and several activation markers and showed markedly enhanced integrin-mediated adhesion to a dermal microvascular endothelial cell line in vitro. Compared with the allostimulated T cells, TILs expressed higher levels of the cutaneous lymphocyte antigen (CLA), the adhesion molecule CD31 and the activation markers CD30 and CD69, but lower levels of several other adhesion and activation molecules. These phenotypic and functional properties of TILs should have complex effects on their migration in vivo. Expression of CLA, the skin homing receptor, may increase migration to melanoma (a skin cancer), whereas integrin activation may cause non-specific binding of TILs to other endothelium. Manipulation of the culture conditions in which TILs are expanded might result in a phenotype that is more conducive to selective tumour homing in vivo.


Carotid intima-media thickness in patients with end-stage renal disease.
Accelerated atherosclerosis and cardiovascular disease are major causes of morbidity and mortality in patients of end-stage renal disease. Carotid intima media thickness is taken as a useful surrogate marker of atherosclerosis. Thirty end-stage renal disease (ESRD) patients were subjected to ultrasonography to study CIMT before the initiation of dialysis. CIMT was found to be higher in ESRD patients than in controls. Levels of a serum marker of oxidative stress were also found to be higher in patients than in the controls. CIMT is an easy, noninvasive, reproducible, and cost-effective investigation in patients with chronic renal failure.


Localization of the neuronal class III beta-tubulin isotype in foci of early neuritogenesis supports divergent neuroblastic differentiation potential in Wilms' tumors.
Wilms' tumors are embryonic neoplasms that have been proposed to originate from the metanephric blastema and are capable of divergent epithelial and mesenchymal differentiation. Neuroepithelial differentiation in these tumors remains controversial. The aim of this study was to examine the phenotypic profile of certain neuronal and glial antigenic determinants in a series of 21 Wilms' tumors. Immunohistochemical studies were performed by using monoclonal antibodies against the neuronal class III beta-tubulin isotype (beta III), the phosphorylated and phosphorylation-independent epitopes of neurofilament protein, and synaptophysin; antisera to gamma-enolase (neuron-specific enolase) glial fibrillary acidic protein, and S100 protein were also used. Foci of neoplastic cells with neurite-like processes that exhibited intense beta III staining were demonstrated in blastemalike areas of three of 21 tumors. In one case, Homer Wright rosettes (stained for beta III) were identified. Areas of abortive neuritic development were also labeled with antibodies to gamma-enolase. No reactivity was obtained in these foci for phosphorylated and phosphorylation-independent epitopes of neurofilament protein, synaptophysin, glial fibrillary acidic protein, and S100 protein. The remainder of the tumors (18 of 21) were negative with the panel of neural markers. Our results indicate that divergent neuroblastic differentiation, evidenced as early neoplastic neuritogenesis, may be present in the blastematous component of Wilms' tumor subsets.


The missing link: improving quality with a chronic disease management intervention for the primary care office.
Bold steps are necessary to improve quality of care for patients with chronic diseases and increase satisfaction of both primary care physicians and patients. Office-based chronic disease management (CDM) workers can achieve these objectives by offering self-management support, maintaining disease registries, and monitoring compliance from the point of care. CDM workers can provide the missing link by connecting patients, primary care physicans, and CDM services sponsored by health plans or in the community. CDM workers should be supported financially by Medicare, Medicaid, and commercial health plans through reimbursements to physicians for units of service, analogous to California's Comprehensive Perinatal Services Program. Care provided by CDM workers should be standardized, and training requirements should be sufficiently flexible to ensure wide dissemination. CDM workers can potentially improve quality while reducing costs for preventable hospitalizations and emergency department visits, but evaluation at multiple levels is recommended.


Ethyl acetate fraction from Angelica sinensis inhibits IL-1β-induced rheumatoid synovial fibroblast proliferation and COX-2, PGE2, and MMPs production.
The root of Angelica sinensis (AS), also known as "Dang-gui," was a popular herbal medicine widely used in the treatment of gynecological diseases in China, Korea, and Japan for a long time. This study aimed to determine the effects of ethyl acetate fraction from Angelica sinensis (EAAS) on the interleukin-1β (IL-1β)-induced proliferation of rheumatoid arthritis synovial fibroblasts (RASFs), and production of matrix metalloproteinases (MMPs), cyclooxygenase (COX) 2, and prostaglandin E2 (PGE2), involved in articular bone and cartilage destruction, by RASFs. RASF proliferation was evaluated with cholecystokinin octapeptide (CCK-8) reagent in the presence of IL-1β with/without EAAS. Expression of MMPs, tissue inhibitor of metalloproteinases-1 (TIMP-1), COXs, PGE2, and intracellular mitogen-activated protein kinase (MAPK) signaling molecules, including p-ERK, p-p38, p-JNK, and NF-κB, were examined using immunoblotting or semi-quantitative reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. EAAS inhibited IL-1β-induced RASF proliferation; MMP-1, MMP-3, and COX-2 mRNA and protein expressions; and PGE2 production. EAAS also inhibits the phosphorylation of ERK-1/2, p38, and JNK, and activation of NF-κB by IL-1β. EAAS might be a new therapeutic modality for rheumatoid arthritis management.


Cancer in relatives of retinoblastoma patients.
Medical histories with emphasis on malignant disease were obtained on 1,269 first and second degree relatives of 93 probands with retinoblastoma and on 671 first and second degree relatives of 50 age-matched control children. The number of nonocular malignancies expected to occur in first and second degree relatives of the probands were calculated using the observed number of neoplasms in control families as the standard. Using a Poisson distribution to evaluate the findings, a statistically significant excess of cancer was found in relatives of the probands with retinoblastoma. Gastrointestinal malignancies were particularly common in this population.


Synthesis and Properties of Subphthalocyanine-Tetracyanobutadiene-Ferrocene Triads.
A series of boron subphthalocyanine-tetracyanobutadiene-ferrocene (SubPc-TCBD-Fc) triads was synthesized by subjecting SubPcs with a ferrocenylethynyl substituent at either the axial or peripheral position to a [2 + 2] cycloaddition reaction with tetracyanoethylene followed by retroelectrocyclization. The ferrocenylethynyl unit was introduced at the axial position (at the boron atom) by a simple aluminum chloride-mediated alkynylation reaction, while functionalization at the SubPc periphery was accomplished by a Sonogashira coupling reaction. The conversion of one alkyne unit into a TCBD unit in combination with the location of the resulting TCBD-Fc moiety was found to have a strong influence on the optical and redox properties, which is ascribed to very different ground-state interactions between the individual donor/acceptor systems. The first electrochemical oxidation could thus be anodically shifted by as much as 0.4 V from the strongest donor molecule (with most unperturbed ferrocene character) to the poorest donor molecule (with strongly perturbed ferrocene character). Six redox states could be reached reversibly for the SubPc-TCBD-Fc triads, -3, -2, -1, 0, + 1, + 2, and for one compound the formation of a tetraanion persistent at the time scale of slow scan voltammetry was observed.


A Mobile Health-Based Management for Patients with Limited Lung Function to Improve Treatment Outcomes and Quality of Life After Minimally Invasive Thoracoscopic Lobectomy.
Limited lung function is an independent risk factor for postoperative respiratory failure in non-small cell lung cancer (NSCLC) patients. In this study, we developed a mobile health-based management for NSCLC patients with limited lung function who were scheduled to receive lobectomy and evaluated its effects on the patient's pulmonary function and quality of life. A total of 60 NSCLC patients scheduled to receive minimally invasive thoracoscopic lobectomy were enrolled and then randomized into the traditional management group and the program management group, with 30 patients per group. Based on the WeChat mini program, a management software for patients with limited lung function was established, including two portals: the patient portal and the nurse one. The pain assessment was performed using the Visual Analog Scale (VAS) scores, the cough assessment using the Leicester Cough Questionnaire, and the quality-of-life assessment using the EORTC QLQ-30 at 1 day before surgery, 1 week, 2 weeks, 1 month, 6 months, or 12 months after surgery. The program management group exhibited an increased PaO2 (96.68 ± 7.92 vs. 87.69 vs. 5.50; P = .018) concomitant with a declined PaCO2 (38.55 ± 2.79 vs. 40.65 ± 2.17; p = 0.034) at 12 months after surgery compared with the traditional management group. The VAS scores showed significant differences at 2 weeks after surgery between the traditional management (median: 2; range: 2-3) and program management (median: 2; range: 1-2) groups (P = .012). The scores of Leicester Cough Questionnaire showed remarkable differences at 12 months after surgery between the traditional management (20.00 ± 1.54) and program management (18.99 ± 2.08) groups (P = .036). The total scores of EORTC QLQ-30 showed notable differences at 12 months after surgery between the traditional management (83.05 ± 14.09) and program management (90.55 ± 11.32) groups (P = .027). The study demonstrated improved pulmonary function and a better quality of life conferred by the mobile health-based management based on WeChat mini program for NSCLC patients with limited lung function and undergoing thoracoscopic lobectomy in a long follow up.


Efficient Reduction of Antibacterial Activity and Cytotoxicity of Fluoroquinolones by Fungal-Mediated N-Oxidation.
Extensive usage of fluoroquinolone antibiotics in livestock results in their occurrence in manure and subsequently in the environment. Fluoroquinolone residues may promote bacterial resistance and are toxic to plants and aquatic organisms. Moreover, fluoroquinolones may enter the food chain through plant uptake, if manure is applied as fertilizer. Thus, the presence of fluoroquinolones in the environment may pose a threat to human and ecological health. In this study, the biotransformation of enrofloxacin, marbofloxacin, and difloxacin by the fungus X. longipes (Xylaria) was investigated. The main metabolites were unequivocally identified as the respective N-oxides by mass spectrometry and nuclear magnetic resonance spectroscopy. Fungal-mediated N-oxidation of fluoroquinolones led to a 77-90% reduction of the initial antibacterial activity. In contrast to their respective parent compounds, N-oxides showed low cytotoxic potential and had a reduced impact on cell proliferation. Thus, biotransformation by X. longipes may represent an effective method for inactivating fluoroquinolones.


Access to skilled care at home during pregnancy and childbirth: Dhaka Bangladesh.
In Bangladesh the majority of childbirth takes place at home by unskilled persons. The objective of this study was to strengthen maternity care services through provision of skilled midwives and to explore people's perception of skilled birth attendants. As part of a quasi-experimental community trial conducted among selected urban area of Dhaka city, a group of skilled midwives were posted in selected urban centers between January to December 2002 to provide obstetric care services and some social mobilization activity in the community. Analysis of skilled midwives self-reported case records and monitory reports of the researchers was done on selected themes. The perception of the skilled midwife was not clear to people. They thought traditional birth attendants who conducts delivery over years were skilled midwives. Preparedness for obstetric emergency and birth planning was non-existent in the families. The skilled midwives were well accepted in the community but discouraged by center authority for home birth, apprehending that it will reduce their client flow and earning. Non-cooperation by other health care providers was common.


A rapid method for assessing the distribution of gold labeling on thin sections.
Particulate gold labeling on ultrathin sections is in widespread use for antigen localization at the EM level. To extend the usefulness of gold labeling technology, we are evaluating different methods for sampling and estimating quantities of gold labeling. Here we present a simple, rapid, and unbiased method for assessing the relative pool sizes of immunogold labeling distributed over different cell compartments. The method uses a sampling approach developed for stereology in which a regular array of microscopic fields or linear scans is positioned randomly on labeled sections. From these readouts, gold particles are counted and assigned to identifiable cell structures to construct a gold labeling frequency distribution of those labeled compartments. Here we use ultrathin cryosections labeled for a range of different proteins and for a signaling lipid. We show by scanning labeled sections at the electron microscope that counting 100-200 particles on each of two grids is sufficient to obtain a reproducible and rapid assessment of the pattern of labeling proportions over 10-16 compartments. If more precise estimates of labeling proportions over individual compartments are required (e.g., to achieve coefficients of error of 10-20%), then 100-200 particles need to be counted over each compartment of interest.


Dispersal in a parasitic worm and its two hosts: consequence for local adaptation.
Characterizing host and parasite population genetic structure and estimating gene flow among populations is essential for understanding coevolutionary interactions between hosts and parasites. We examined the population genetic structure of the trematode Schistosoma mansoni and its two host species (the definitive host Rattus rattus and the intermediate host Biomphalaria glabrata) using microsatellite markers. Parasites were sampled from rats. The study was conducted in five sites of the Guadeloupe Island, Lesser Antilles. Mollusks display a pattern of isolation by distance whereas such a pattern is not found neither in schistosomes nor in rats. The comparison of the distribution of genetic variability in S. mansoni and its two host species strongly suggests that migration of parasites is principally determined by that of the vertebrate host in the marshy focus of Guadeloupe. However, the comparison between genetic differentiation values in schistosomes and rats suggests that the efficacy of the schistosome rat-mediated dispersal between transmission sites is lower than expected given the prevalence, parasitic load and migration rate of rats among sites. This could notably suggest that rat migration rate could be negatively correlated to the age or the infection status of individuals. Models made about the evolution of local adaptation in function of the dispersal rates of hosts and parasites suggest that rats and mollusks should be locally adapted to their parasites.


Enhancing knowledge of Diabetes self-management and quality of life in people with Diabetes Mellitus by using Guru Diabetes Apps-based health coaching.
Virtual diabetes coaching through applications has the potential to improve self-management in people with Type 2 Diabetes Mellitus. However, applications designed for Asian's diabetic have never been studied. Therefore, our study aimed to identify the effect of Guru Diabetes Apps-Based Health Coaching on the level of glycemic hemoglobin, knowledge, and quality of life in Indonesians, as one of Asian ethnicities. This study used an experimental design. Participants with T2DM were recruited from three primary care health centers. The intervention included 3 months of virtual health coaching through the Guru Diabetes application. The daily monitoring of glycemic blood glucose, blood pressure, body weight, nutrition intake, and activity provided were used as baseline to conduct virtual in-person coaching. The glycemic hemoglobin level, knowledge level, and quality of life were assessed at baseline and post-enrollment. A multilevel modeling approach was used for statistical analysis. A total of 66 respondents were enrolled, and 62 respondents (93.93%) remained in the study at 3 months. After completing the intervention, the HbA1c level decreased in all groups, however it did not differ among the groups. On the other hand, the results indicate significant improvement in the average diabetes self-management knowledge score (pre = 14.97 vs post = 19.07, p < 0.05) and quality of life score (pre = 54.34 vs post = 60.28, p < 0.05), with significant difference among the groups (p < 0.05). Participants in the Android Application-Based Health Coaching experienced a significant improvement in diabetes self-management knowledge and quality of life scores at 3 months. In conclusion, the Guru Diabetes Apps-Based Health Coaching has the potential to support people with T2DM in performing diabetes self-management at home.


Interpolation of multidimensional diabatic potential energy matrices.
A method for constructing diabatic potential energy matrices by interpolation of ab initio quantum chemistry data is described and tested. This approach is applicable to any number of interacting electronic states, and relies on a formalism and a computational procedure that are more general than those presented previously for the case of two electronic states. The method is tested against an analytic model for three interacting electronic states of NH(3) (+).


Sperm morphology assessment: diagnostic potential and comparative analysis of strict or WHO criteria in a fertile and a subfertile population.
This prospective study compared the diagnostic and predictive potential of sperm morphology assessments in a fertile vs. a subfertile population, evaluated in three different laboratories. The fertile population included 144 men who had recently fertilized their partners. As subfertile controls, 136 men with a history of subfertility for more than 12 months were used. All semen samples (280) were evaluated in three different centres in a blind fashion, without any patient information. The evaluation of sperm morphology was performed according to the criteria normally used in the different laboratories: WHO (1992) criteria for laboratory A, and Tygerberg strict criteria for laboratories B and C. Using ROC analysis, the predictive power of sperm morphology turned out to be different in the three laboratories (area under ROC curve: 69% for lab A, 72% for lab B and 78% for lab C). Using percentile 10 of the fertile population as the cut-off value for normality, we obtained the following results: 2, 1 and 5% for laboratories A, B and C, respectively. Using ROC analysis cut-off values with optimal specificity and sensitivity were 6, 1 and 10%, respectively. Although our data highlight a reasonable predictive power of sperm morphology in centres using different or the same criteria, cut-off values for normality were different, even when the same criteria were applied. These results stress the importance of standardization in sperm morphology evaluation and the need for examining a reference population in estimating the real threshold value in different laboratories.


Fidelity of implementation to a care team redesign and improved outcomes of diabetes care.
We assessed fidelity of implementation (FOI) to the intended features of a primary care team redesign that integrated registered nurse care managers and patient health coaches onto existing care teams. The relation of FOI ranking and improvements in intermediate outcomes of diabetes care was examined. We assessed FOI by interviewing frontline primary care team members (n = 20). We explored the relation of FOI and outcomes of diabetes care (n = 10 206 patients) over a 3-year period (2010-12). Multilevel, multivariate regression estimated the relation of FOI and improvements in outcomes of diabetes care. Five primary care practices in greater Los Angeles, CA, USA. Ten thousand, two hundred and six adult patients with diabetes; 20 frontline primary care clinicians and staff. Blood pressure, hemoglobin A1c (HbA1c) and low-density lipoprotein cholesterol (LDL-C) control among adult diabetic patients. All practices improved diabetic patients' LDL-C control over time. In adjusted analyses, the practice with the highest FOI achieved the largest improvement in blood pressure and HbA1c control among diabetic patients. In contrast, the practice with the lowest FOI had the least improvements in blood pressure, HbA1c and LDL-C control. FOI was an inconsistent predictor of intermediate outcomes of diabetes care for other practices. FOI assessment can be useful for identifying low FOI to a redesign so that technical assistance and resources can be provided to improve team functioning and patient outcomes. High FOI can enable greater improvements in patient outcomes in the context of primary care practice redesign.


What are the Particularities of Splenic Surgery in Cirrhotic Patients?
The aim of this work was to review the entire literature on splenic surgery in cirrhotic patients in order to best define the surgical indications and their management specifics. A review of the international literature published between January 1995 and August 2015 was thus carried out.


A study on Candida biofilm growth characteristics and its susceptibility to aureobasidin A.
Biofilm is known to contribute to the antifungal resistance of Candida yeasts. Aureobasidin A (AbA), a cyclic depsipeptide targeting fungal sphingolipid biosynthesis, has been shown to be effective against several Candida species. The aim of this study was to investigate Candida biofilm growth morphology, its biomass, metabolic activity, and to determine the effects of AbA on the biofilm growth. The biofilm forming ability of several clinical isolates of different Candida species from our culture collection was determined using established methods (crystal violet and XTT assays). The determination of AbA planktonic and biofilm MICs was performed based on a micro-broth dilution method. The anti-biofilm effect of AbA on Candida albicans was examined using field emission scanning electron microscope (FESEM) analysis. A total of 35 (29.7%) of 118 Candida isolates were regarded as biofilm producers in this study. Candida parapsilosis was the largest producer, followed by Candida tropicalis and C. albicans. Two morphological variants of biofilms were identified in our isolates, with 48.6% of the isolates showing mainly yeast and pseudohyphae-like structures, while the remaining ones were predominantly filamentous forms. The biofilm producers were divided into two populations (low and high), based on the ability in producing biomass and their metabolic activity. Candida isolates with filamentous growth, higher biomass and metabolic activity showed lower AbA MIC50 (at least fourfold), compared to those exhibiting yeast morphology, and lower biomass and metabolic activity. The observation of filament detachment and the almost complete removal of biofilm from AbA-treated C. albicans biofilm in FESEM analysis suggests an anti-biofilm effect of AbA. The variability in the growth characteristics of Candida biofilm cultures affects susceptibility to AbA, with higher susceptibility noted in biofilm cultures exhibiting filamentous form and high biomass/metabolic activity.


Cell surface phenotyping of megakaryoblasts.
Surface phenotypic characterization of megakaryoblasts, identified by platelet peroxidase activity, was investigated in four patients who showed increased proliferation of megakaryoblasts: one patient with typical features of acute leukemia, one presenting with acute myelofibrosis, and two with Down's syndrome in whom blasts disappeared spontaneously (transient abnormal myelopoiesis, TAM). MY10 and/or MY9 antigens were expressed on the surface of some megakaryoblasts, but MY7, and MY4, antigens specific to granulocytic or monocytic cells, were not. Some megakaryoblasts were positive for only anti-HLA-DR antibodies. It was speculated that, during the differentiation of the megakaryocytic lineage, MY9 antigen appears transiently on the surface of megakaryoblasts that have lost HLA-DR antigens and have gained the glycoprotein IIb/IIIa antigen. This study also demonstrated that the proliferating blasts in some patients with TAM were mainly megakaryoblasts and suggested that the target cells in TAM are CFU-GEMM.


COVID-19: A learning moment for patients and health professionals.
A hospital physician from Northern Italy describes his own experience as caregiver of COVID-19 patients and as a patient himself who required treatment in urgent care. From this experience he learned that an untapped reserve of human solidarity exists in a team of caregivers in the midst of a crisis where they find an unsuspected shared energy. He never would have believed to be able to work long hours patiently and without sleep until he was challenged by the demands of the COVID crisis. As such, he discovers that to be effective the team leader should lead by example rather than by commandments! His experience as a patient allowed him to face his own mortality, to learn that healing rather than cure is the ultimate goal of caregiving.


Breast cancer risk-factor and screening awareness among women nurses and teachers in Amman, Jordan.
Breast cancer awareness studies of women in Jordan do not exist. This study used data from 163 nurses and 178 teachers surveyed in Amman to determine 2 dimensions of breast cancer awareness: general breast cancer awareness, defined as knowledge of risk factors associated with the disease and breast cancer screening awareness, defined as knowledge of breast self-examination and mammography. The survey instrument was based on 2 previously validated knowledge-based questionnaires in the literature (Breast Cancer Knowledge Test and the Comprehensive Breast Cancer Questionnaire). Analysis of covariance indicated that family history was associated with general breast cancer awareness. Profession, age, and family history significantly influenced breast cancer screening awareness. The average percentage of correct responses to general breast cancer awareness was adjusted for select covariates (adjusted means). The adjusted mean general awareness score for nurses was not significantly different from that of teachers (P =.8470). Nurses were more aware than teachers of the importance of breast cancer screening and its techniques. The adjusted mean screening awareness score for nurses was 88.3%, compared with 73.1% for teachers (P <.0001). These results provide important information about the level of breast cancer awareness among women nurses and teachers in Jordan and may be useful for developing future prevention and screening education programs.


Induction of a phenobarbital-inducible form of cytochrome P-450 in rat liver microsomes by 1,1-di(p-chlorophenyl)-2,2-dichloroethylene.
When 1,1-di(p-chlorophenyl)-2,2-dichloroethylene (DDE) (100 mg/kg body weight) was injected into rats, the benzphetamine N-demethylation and 7-ethoxycoumarin O-deethylation activities of liver microsomes increased by 7-fold and 3-fold, respectively, at 7 days after the injection, whereas the benzo(a)pyrene 3-hydroxylation activity did not increase. The content of cytochrome P-450 in the microsomes increased 2.5-fold at 7 days. By the use of the antibodies to a phenobarbital (PB)-inducible form (P-450(PB-1)) and a 3-methylcholanthrene (MC)-inducible form (P-450(MC-1)) of cytochrome P-450, the contents of P-450(PB-1) and P-450(MC-1) in the liver microsomes of DDE-treated rats were measured. The form of cytochrome P-450 immunoprecipitable with anti-P-450(PB-1) antibodies increased by 10-fold at 7 days. A major component of cytochrome P-450 in the liver microsomes of DDE-treated rats, which was tentatively designated P-450(DDE), was purified. P-450 (DDE) was compared with P-450(PB-1), and they were found to be indistinguishable by the following criteria: 1) chromatographic behavior on aminooctyl-Sepharose 4B, hydroxyapatite, and DEAE-cellulose columns, 2) minimum molecular weight determined by SDS-polyacrylamide gel electrophoresis, 3) spectral properties, 4) immunoreactivity, 5) amino acid composition, 6) peptide mapping, 7) NH2-terminal amino acid sequence, 8) catalytic activities. We concluded that DDE and PB induce an identical form of P-450 in rat liver microsomes, although DDE is apparently very different from PB in chemical structure.


Evaluation of Parotid Salivary Glucose Level for Clinical Diagnosis and Monitoring Type 2 Diabetes Mellitus Patients.
Background. To investigate the relationships among blood glucose, mixed saliva glucose, and parotid glucose in type 2 diabetes patients and to evaluate the diagnostic and monitoring value of salivary gland glucose in patients with type 2 diabetes (type 2DM). Material and Methods. Thirty patients with type 2DM and 30 healthy age- and sex-matched individuals were included in this study. Glucose levels in unstimulated mixed saliva and in unstimulated parotid saliva were measured by the glucose oxidase peroxidase method. Results. The blood glucose and parotid salivary glucose levels in type 2DM patients were significantly higher than those in the controls (P < 0.05). The blood glucose, parotid salivary glucose, and mixed salivary glucose were 7.46 ± 1.44 mmol/L, 0.18 ± 0.19 mmol/L, and 3.17 × 10-2 ± 2.84 × 10-2 mmol/L, respectively, in the type 2DM group; the corresponding glucose levels in the control group were 5.56 ± 0.71 mmol/L, 7.70 × 10-2 ± 6.02 × 10-2 mmol/L, and 3.47 × 10-2 ± 2.79 × 10-2 mmol/L. The parotid salivary and blood glucose levels in type 2DM patients were strongly correlated; the linear regression equation for blood glucose and parotid salivary glucose was Y = 6.267X + 6.360, with r = 0.810. However, mixed salivary glucose levels were not significantly different in the type 2 diabetes group compared with the control group. Conclusion. Our results suggest that parotid salivary glucose has potential as a biomarker to monitor type 2DM and as a painless, noninvasive method for the management of type 2DM.


Fractal analysis of the vascular tree in the human retina.
The retinal circulation of the normal human retinal vasculature is statistically self-similar and fractal. Studies from several groups present strong evidence that the fractal dimension of the blood vessels in the normal human retina is approximately 1.7. This is the same fractal dimension that is found for a diffusion-limited growth process, and it may have implications for the embryological development of the retinal vascular system. The methods of determining the fractal dimension for branching trees are reviewed together with proposed models for the optimal formation (Murray Principle) of the branching vascular tree in the human retina and the branching pattern of the human bronchial tree. The limitations of fractal analysis of branching biological structures are evaluated. Understanding the design principles of branching vascular systems and the human bronchial tree may find applications in tissue and organ engineering, i.e., bioartificial organs for both liver and kidney.


Adverse reactions to OsteoSet bone graft substitute, the incidence in a consecutive series.
The purpose of this study was to identify factors that may be associated with the development of a sterile inflammatory reaction to OsteoSet (Wright Medical Technology) bone graft substitute. We report on a series of 58 consecutive cases of patients with benign bone lesions who developed an adverse reaction to Osteoset. All procedures were performed by a single orthopaedic oncology surgeon. Patients who exhibited an adverse reaction to OsteoSet and those who did not were analyzed for statistical differences with respect to age, sex, volume of graft material used, size of pellet, use of alcohol washes, mixing of OsteoSet with allograft, medications, drug allergies, and smoking history. Comparisons were made between the reaction group and the non-reaction group with respect to the these variables. Eight definite (13.8%) and 3 probable (5.2%) reactions were identified. Symptoms developed an average of 38.9 days after the operation. There was no statistically significant difference (p>0.05) in the volume of Osteoset used, size of pellet, use of alcohol washes, mixing of OsteoSet with allograft, patient age, sex, drug allergies, or smoking history. Reported use of at least one medication was statistically significant between groups when probable reactions were included with the non-reaction group (p < 0.05), but not when probables were analyzed with the definite reaction group ( p < 0.10). A review of our patients' medication lists did not reveal any single or combination of medications likely to be responsible for this difference. Type and location of tumor also did not appear to differ significantly between the two groups. Although we report a 13.8%-19.0% incidence of adverse reaction to OsteoSet, we are unable to identify any statistically significant factors that might predict the development of a reaction. Fortunately, the reaction is essentially self limited and fairly benign requiring only anti-inflammatories and observation.


Cultural perspectives on pain management.
Bedouin women tend to remain quiet and expressionless while giving birth despite reporting high levels of pain and fear (Harrison 1991). Culture undoubtedly influences pain perception and expression but there are dangers in making assumptions about particular groups. This article explores the underlying research and the complex clinical picture highlighting the need for cultural awareness--but ultimately we must care for the individual person in pain.


Drug Targets for Cardiovascular-Safe Anti-Inflammatory: In Silico Rational Drug Studies.
Cyclooxygenase-2 (COX-2) plays an important role in memory consolidation and synaptic activity, the most fundamental functions of the brain. It converts arachidonic acid to prostaglandin endoperoxide H2. In contrast, if over-expressed, it causes inflammation in response to cytokine, pro-inflammatory molecule, and growth factor. Anti-inflammatory agents, by allosteric or competitive inhibition of COX-2, alleviate the symptoms of inflammation. Coxib family drugs, particularly celecoxib, are the most famous anti-inflammatory agents available in the market showing significant inhibitory effect on COX-2 activity. Due to high cardiovascular risk of this drug group, recent researches are focused on the investigation of new safer drugs for anti-inflammatory diseases. Natural compounds, particularly, phytochemicals are found to be good candidates for drug designing and discovery. In the present study, we performed in silico studies to quantitatively scrutinize the molecular interaction of curcumin and its structural analogs with COX-2, COX-1, FXa and integrin αIIbβIII to investigate their therapeutic potential as a cardiovascular-safe anti-inflammatory medicine (CVSAIM). The results of both ADMET and docking study indicated that out of all the 39 compounds studied, caffeic acid had remarkable interaction with proteins involved in inflammatory response. It was also found to inhibit the proteins that are involved in thrombosis, thereby, having the potential to be developed as therapeutic agent.


Acute encephalopathy with bilateral thalamotegmental involvement and a benign course: a case report from Brazil.
This rare encephalopathy that generally affects children is preceded by a respiratory infection and fever associated with convulsions and may progress to coma. Outcome is catastrophic in most cases. This case report describes a Brazilian child of African descent with fever, cephalea and bilateral amaurosis, who evolved to coma with pyramidal signs and associated convulsions. MRI showed diffuse, symmetrical lesions in the thalamotegmental region and brainstem. Following administration of methylprednisolone, the clinical condition of the patient improved and the brain lesions regressed, leaving the child with no current neurological deficits. This was a case of acute postinfectious encephalopathy, involving various brain structures. Outcome was favourable with no sequelae following therapy. This case was atypical due to the bilateral visual involvement and extensive encephalic lesions in a child of African descent with no neurological sequelae following therapy. No other similar cases have been reported in the literature.


Efficacy and safety of trastuzumab combined with chemotherapy for first-line treatment and beyond progression of HER2-overexpressing advanced breast cancer.
To observe the efficacy and safety of trastuzumab combined with chemotherapy in patients with human epidermal growth factor receptor 2 (HER2)-overexpressing advanced breast cancer. A total of 90 patients with HER2-overexpressing advanced breast cancer were enrolled in this study. All patients were diagnosed with ductal invasive breast cancer by pathological analysis, and were aged between 31-73 years with a median of 51 years. HER2-positivity was defined as 3(+) staining in immunochemistry or amplification of fluorescence in situ hybridization (FISH, ratio ≥2.0). Trastuzumab was administered in combination with chemotherapy as first-line treatment and beyond progression as a secondline, third-line, and above treatment in 90, 34, 14, and 6 patients, respectively. The chemotherapy regimen was given according to normal clinical practice. The response rate was evaluated every two cycles, and the primary endpoints were progression-free survival (PFS) and overall survival (OS). Survival curves were estimated by using Kaplan-Meier graphs and were compared by using log-rank test statistics. Multivariate analysis was done using Cox's proportional hazards regression model, and the level of significance was P<0.05. All 90 patients received at least one dose of trastuzumab, and efficacy could be evaluated in 85 patients. The median follow-up was 50 months. In total, 72 (80.00%) patients had visceral metastasis, and 43 (47.78%) patients had progressed after one or more extensive chemotherapy regimens for metastatic diseases. The median PFS for first-line trastuzumab was 10 months (range, 2-59 months), and the median OS after metastasis or initially local advanced disease was 22 months (range, 2-116 months). Trastuzumab combined with chemotherapy was active and well-tolerated as a first-line treatment and even beyond progression in HER2-overexpressing advanced breast cancer as a second-line or third-line treatment. However, its efficacy is certainly less beyond this point.


[Rheumatic diseases as risk factors for cardiovascular disease].
Cardiovascular disease is the leading cause of death in patients with rheumatoid arthritis (RA), ankylosing spondylitis (AS) and systemic lupus erythematosus (SLE). In addition to mortality, cardiovascular morbidity is also markedly increased in these patients, compared with the general population. The increased cardiovascular risk can be explained only partially by an increased prevalence of classical risk factors for cardiovascular disease; it also appears to be related to inflammation. Prospective intervention trials aimed at the modification of cardiovascular risk factors are needed to determine the impact of cardiovascular risk reduction in patients with rheumatic disease. In addition to SLE, RA and AS should be acknowledged as new risk factors for cardiovascular disease.


Opioid agonist treatment reduces losses in quality of life and quality-adjusted life expectancy in heroin users: Evidence from real world data.
This study estimated the long-term changes of opioid agonist treatment (OAT) in quality of life (QOL) and quantified the quality-adjusted life years (QALY) from the loss of quality-adjusted life expectancy (QALE) in heroin users. A total of 1283 heroin users stratified by OAT were linked to the National Mortality Registry for 8 years (2006-2014) to obtain survival functions, which were extrapolated to lifetime by applying a rolling extrapolation algorithm to survival ratio between the sub-cohorts and age- and sex-matched referents simulated from vital statistics of Taiwan. We performed cross-sectional measurement of EQ-5D on 349 participants, including those with a valid state of OAT or non-OAT plus newly recruited consecutive patients, during 2015-2017 for utility values, while the QOL of referents were abstracted from the 2009 National Health Interview Survey. The QALE was calculated by summing the products of the mean QOL and survival rate throughout life. The QALE difference between the cohort and corresponding referents was the loss-of-QALE. QOL of the OAT group was significantly better than that of the non-OAT group in every domain of the EQ-5D, which was quantified to be 0.23 for utility after controlling for other variables. After extrapolation to 70 years, the estimated QALE and loss-of-QALE were 17.8 and 18.2 QALY for OAT subjects, respectively, while those of the non-OAT group were 9.2 and 27.9 QALY. Receiving OAT could reduce QALE lost by 9.7 QALYs compared with non-OAT after accounting for QOL differences along time and different age and sex distributions.


An indocyanine green-based liquid biopsy test for circulating tumor cells for pediatric liver cancer.
Hepatoblastoma (HB) and hepatocellular carcinoma (HCC) are the most common malignant hepatocellular tumors seen in children. The aim of this work was to develop a liquid biopsy test for circulating tumor cells (CTCs) for these tumors that would be less invasive and provide information about the real-time state of tumors in response to therapies. For this test, we utilized indocyanine green (ICG), a far-red fluorescent dye that is used clinically to identify malignant liver cells in the body during surgery. We assessed ICG accumulation in cell lines with fluorescence microscopy and flow cytometry. For our CTC test, we developed a panel of liver tumor-specific markers, ICG, Glypican-3 (GPC3), and DAPI and tested this panel with cell lines and non-cancer control blood samples. We then used this panel to analyze whole blood samples for CTC burden with a cohort of 14 HB and HCC patients and correlated with patient characteristics and outcomes. We showed that ICG accumulation is specific to liver cancer cells, compared to non-malignant liver cells, non-liver solid tumor cells, and non-malignant cells and can be used to identify liver tumor cells in a mixed population of cells. Experiments with the ICG/GPC3/DAPI panel showed that it specifically tagged malignant liver cells. With patient samples, we found that CTC burden from sequential blood samples from the same patients mirrored the patients' responses to therapy. Our novel ICG-based liquid biopsy test for CTCs can be used to specifically count CTCs in the blood of pediatric liver cancer patients. This manuscript represents the first report of circulating tumor cells in the blood of pediatric liver cancer patients. The novel and innovative assay for CTCs shown in this paper will facilitate future work examining the relationship between CTC numbers and patient outcomes, forming the foundation for incorporation of liquid biopsy into routine clinical care for these patients. Overview of novel liquid biopsy test for circulating tumor cells for pediatric liver cancer. Figure made with Biorender.


Organizational psychosocial factors and deployment-related exposure concerns in Afghanistan/Iraq War veterans.
Environmental exposure concerns are associated with adverse health outcomes in soldiers deployed to South West Asia. There is little data on factors associated with the reporting of exposure concerns. We explored the relationship between deployment-related preparedness/support and exposure concerns. Retrospective chart review of 489 Afghanistan/Iraq veterans evaluated at a Veterans Affairs tertiary center for postdeployment health. Virtually all subjects were concerned about environmental exposure(s). There were no significant demographic differences in exposure concerns, preparedness/support variables, or both. Preparedness/support correlated inversely with exposure concerns. Mental health function mediated the relationship between preparedness/support and exposure concerns. Deployment-related preparedness/support is associated with exposure concerns and mental health functioning. Definitive studies will provide data and insight on how the military may better prepare/support soldiers to optimize their resilience and reduce deployment-related exposure concerns.


Static cardiomyoplasty with synthetic elastic net suppresses ventricular dilatation and dysfunction after myocardial infarction in the rat: a chronic study.
Although static cardiomyoplasty prevents the left ventricle (LV) from dilatation, it may interfere with diastolic relaxation, or cause restriction. We developed a synthetic net with dual elasticity and tested its effect late after myocardial infarction in the rat. LV pressure-volume relationships (PVR) were successively analyzed before, after intravenous volume load, and 10 minutes after occlusion of the left anterior descending artery. Rats were then randomized into groups receiving synthetic net wrapping around the heart (NET+, n = 8) and only partially behind LV (NET-, n = 9), and they underwent the same PVR studies 6 weeks later. End-diastolic and end-systolic PVR were defined, and LV size and function were compared under standardized loading conditions. Although there was no difference in Day 0, increase in LV end-diastolic and end-systolic volumes were significantly attenuated in NET+ rats 6 weeks later when there was a significant correlation between LV volumes by PVR estimation and actual measurements, with significant differences in both measures between the groups: NET+ < NET-. The presence or absence of net did not show restrictive hemodynamics under acute volume load. Static cardiomyoplasty using a synthetic elastic net significantly attenuated LV dilatation and dysfunction without restriction late after myocardial infarction in the rat.


Bioinformatics analysis of the structural and evolutionary characteristics for toll-like receptor 15.
Toll-like receptors (TLRs) play important role in the innate immune system. TLR15 is reported to have a unique role in defense against pathogens, but its structural and evolution characterizations are still poorly understood. In this study, we identified 57 completed TLR15 genes from avian and reptilian genomes. TLR15 clustered into an individual clade and was closely related to family 1 on the phylogenetic tree. Unlike the TLRs in family 1 with the broken asparagine ladders in the middle, TLR15 ectodomain had an intact asparagine ladder that is critical to maintain the overall shape of ectodomain. The conservation analysis found that TLR15 ectodomain had a highly evolutionarily conserved region on the convex surface of LRR11 module, which is probably involved in TLR15 activation process. Furthermore, the protein-protein docking analysis indicated that TLR15 TIR domains have the potential to form homodimers, the predicted interaction interface of TIR dimer was formed mainly by residues from the BB-loops and αC-helixes. Although TLR15 mainly underwent purifying selection, we detected 27 sites under positive selection for TLR15, 24 of which are located on its ectodomain. Our observations suggest the structural features of TLR15 which may be relevant to its function, but which requires further experimental validation.


Overexpression of LASS2 inhibits proliferation and causes G0/G1 cell cycle arrest in papillary thyroid cancer.
The aim of this study was to investigate the role of LAG1 longevity-assurance homologue 2 (LASS2) in papillary thyroid cancer (PTC). Immunohistochemistry staining was conducted to explore the expression levels of LASS2 in PTC tissues and adjacent normal thyroid tissues and nodular goiter tissues. Western blotting and RT-qPCR were performed to explore the expression levels of LASS2 in three PTC cell lines (TPC-1, K1, BCPAP). An Adv-LASS2-GFP recombinant adenovirus vector was constructed and transduced into BCPAP cells. Then CCK-8 assay, colony formation assay, cell cycle distribution, and apoptosis were performed. Western blotting was used to examine the expression of p21, cyclin D1, cyclin-dependent kinase 4, p53 and p-p53. LASS2 was downregulated in PTC tissues compared with adjacent thyroid tissues or nodular goiter tissues. In addition, the expression of LASS2 was found to be associated with TNM stage and lymph node metastasis. BCPAP cells expressed the lowest LASS2 compared to TPC-1 cells or K1 cells. Overexpression of LASS2 significantly inhibited proliferation, promoted apoptosis and caused G0/G1 cell cycle arrest in BCPAP cells. Furthermore, overexpression of LASS2 significantly increased the expression of p21, inhibited the expression of cyclin D1 and cyclin-dependent kinase 4, and increased the expression of p-p53, but did not effect the expression of p53 in BCPAP cells. Our findings indicate that overexpression of LASS2 inhibits PTC cell proliferation, promotes apoptosis and causes G0/G1 cell cycle arrest via a p53-dependent pathway. Thus, LASS2 may serve as a novel biomarker in PTC.


[Connotation of channel and its relation with nerve].
Originated for the recognition on the anatomy and physiology of "vessel", the theory of channel treats the running pathway of the vessel as jing mai (channel vessel), with its transverse branches as mai (vessel), thus the idea of jing luo and jing mai was gradually formed and the connotation of "channel (jing luo)" is with the etymological connotation of function of the nerve. The challenge to channel theory comes from the introduction of western medicine, or in other words, people interpret the communication among the body surface and the upper and lower parts of the surface, the special connection between the body surface and the internal organs, the adjusting and reaction function. During the introduction of western anatomy-physiology in the late Ming dynasty or the eastward introduction of western medical science at the late Qing dynasty, the terms of traditional Chinese medicine (TCM) connected with channel functions, including "minute tendon (xi jin)", "connection (xi)" had been rendered as the Chinese translation of "nerve". Thus, the language transforming basis between channel and nerve function was laid down at the first stage of confluence between TCM and western medicine.


Duration of fluoroscopic-guided spine interventions and radiation exposure is increased in overweight patients.
The impact of patient body mass index (BMI) on image-guided spine interventions remains unknown. Higher BMI is known to complicate the acquisition of radiographic images. Therefore it can be hypothesized that the patient's body habitus can influence the delivery of a spinal injection. To quantify the impact of patient BMI on the length of fluoroscopy and procedure times during spine interventions. Secondary analysis of 2 prospective observational studies. All injections were performed in an outpatient university setting. A total of 209 patients in whom spine injections were performed (99 women), with a mean age of 54.6 years. The fluoroscopy times for 202 participants and total procedure times for 137 participants were recorded. Additional participant characteristics, including age, gender, BMI, and actual procedures performed, also were collected. Analysis of covariance and linear and nonlinear model analysis were performed to assess the effect of BMI on fluoroscopy and procedure times. Fluoroscopy time and procedure duration times. Participants had a mean age of 54.6 years, 51% were men, and 77% (n = 155) were overweight (BMI ≥25). Participants received the following interventions: 40 zygapophyseal joint injections, 33 medial branch nerve blocks, 113 transforaminal epidural injections, and 16 combined zygapophyseal joint injections and epidural injections. Gender, procedure number, and procedure type did not differ between groups. The overweight group demonstrated a 30% increase in mean fluoroscopy time and a 35% increase in mean procedure time. Controlling for other variables, we found that differences in fluoroscopy time and procedure time were significant (P = .032 and P = .031, respectively) between the 2 groups. Significantly prolonged procedure time and fluoroscopy time in overweight patients increase the risks associated with spine interventions, not only to the patients but also to the operating room staff exposed to ionizing radiation.


Use of the short form 36 (SF36) for health status measurement in rheumatoid arthritis.
The patient-administered Health Assessment Questionnaire (HAQ) is widely used in rheumatology studies. Another health quality assessment technique commonly used for other non-rheumatological conditions is the 'Short Form 36' (SF36). This has questions designed to assess eight aspects of health ranging from physical limitations to general perceptions of vitality and mental well-being. This study presents information on the health status of 137 patients with rheumatoid arthritis (RA) assessed by both the SF36 and HAQ. Summary statistics are given for the elements of the SF36 according to age, gender, disease measures of RA and the presence of co-morbidity. There were significant associations between the physical functioning score of the SF36 and the HAQ score. with other measures of disease activity and severity, and with co-morbidity, although there was considerable inter-patient variability. These findings suggest that future applications of health status questionnaires are possible.


Serum levels of excitatory amino acids, serine, glycine, histidine, threonine, taurine, alanine and arginine in treatment-resistant depression: modulation by treatment with antidepressants and prediction of clinical responsivity.
Previous research has revealed that major depression is accompanied by disorders in excitatory amino acids, e.g. glutamate and aspartate, and alterations in serum levels of other amino acids, e.g. serine, glycine and taurine. The aim of the present study was to examine serum levels of aspartate, asparagine, glutamate, glutamine, serine, glycine, threonine, histidine, alanine, taurine and arginine in major depression patients with treatment-resistant depression (TRD). No significant differences in the serum concentrations of any of the above amino acids could be found between patients with and without TRD and normal controls. Non-responders to treatment with antidepressants during a period of 5 weeks were characterized by significantly lower serum levels of aspartate, asparagine, serine, threonine and taurine. A 5-week period of treatment with antidepressants significantly reduced the serum levels of aspartate, glutamate and taurine, and significantly increased the serum concentrations of glutamine. The results suggest that alterations in serum levels of aspartate, asparagine, serine, threonine and taurine may predict the subsequent response to treatment with antidepressants, and that the latter may modulate serum levels of excitatory amino acids and taurine.


Long Non-Coding RNA KCNQ1OT1 Promotes Cataractogenesis via miR-214 and Activation of the Caspase-1 Pathway.
KCNQ1OT1 regulates the expression of tissue-specific imprinted genes within the Kcnq1 domain. Imprinted genes are positive regulators of apoptosis, one of the forms of cell death related to cataract formation, and thus may provide novel therapeutic targets for cataract treatment. Here, we studied the role of non-coding RNAs(ncRNA) in cataract formation. Human lens epithelium cells (HLECs) were treated with H2O2, and the expression of KCNQ1OT1 and miR-214 was detected by qRT-PCR. The expression of caspase-1 was detected using qRT-PCR, western blot and immunostaining. To confirm our findings in cell cultures, we analysed KCNQ1OT1, miR-214, and caspase-1 expression in lens anterior capsules of both cataract patients and normal controls by qRT-PCR and western blot analysis. We found that the expression of KCNQ1OT1 was increased in both human cataract lens anterior capsular samples and SRA01/04 cell lines treated with H2O2. Knockdown of KCNQ1OT1 expression significantly suppressed H2O2-induced SRA01/04 cell pyroptosis in vitro, which is the critical step in cataract formation. The expression of microRNA-214 (miR-214) was also decreased in cataract lens anterior capsular tissues and H2O2-induced SRA01/04 cell lines. Knockdown of KCNQ1OT1 significantly increased the expression of miR-214. We demonstrated for the first time that caspase-1 is a functional downstream target of miR-214, and knockdown of KCNQ1OT1 reduced the expression of caspase-1. These results provide evidence that the KCNQ1OT1-miR-214-caspase-1 regulatory network is a novel mechanism for promoting cataract formation.


Neonatal and adult gonadal hormone manipulations enhance morphine analgesia elicited from the ventrolateral periaqueductal gray in female rats.
Male rodents displayed greater magnitudes of analgesia following systemic, ventricular, and intracerebral administration of mu-opioid receptor agonists than female rodents. Whereas neonatal castration of male rat pups produced reductions in systemic and central morphine analgesia as adults, neonatal androgenization of female rat pups treated with testosterone propionate (TP) displayed enhancements in systemic and central morphine analgesia as adults. Adult gonadectomy minimally affected mu-opioid analgesia, except if less potent mu agonists were employed, or if morphine was directly administered into the ventrolateral periaqueductal gray (vlPAG). Adult ovariectomy failed to appreciably alter the enhanced analgesia following systemic morphine in female rats with neonatal androgenization. Because the vlPAG elicited morphine analgesia that was sensitive to both neonatal and adult gonadal hormone manipulations, the present study examined morphine analgesia elicited from the vlPAG in female rats receiving neonatal treatment with TP or vehicle and subsequently exposed to adult ovariectomy or sham surgery as well as intact male rats. Intact male rats displayed significantly greater magnitudes and potencies in vlPAG morphine analgesia than female rats receiving neonatal treatment with either vehicle or TP. In turn, neonatal androgenization significantly enhanced vlPAG morphine analgesia relative to neonatal vehicle treatment in females. Adult ovariectomy significantly enhanced the magnitude of vlPAG morphine analgesia in female rats receiving neonatal treatment with either vehicle or TP. This demonstrates a strong interaction between neonatal and adult gonadal hormone manipulations in the mediation of vlPAG morphine analgesia in female rats.


Receptor mechanisms underlying the CNS effects of cannabinoids: CB1 receptor and beyond.
Cannabis legalization continues to progress in many US states and other countries. Δ9-tetrahydrocannabinol (Δ9-THC) is the major psychoactive constituent in cannabis underlying both its abuse potential and the majority of therapeutic applications. However, the neural mechanisms underlying cannabis action are not fully understood. In this chapter, we first review recent progress in cannabinoid receptor research, and then examine the acute CNS effects of Δ9-THC or other cannabinoids (WIN55212-2) with a focus on their receptor mechanisms. In experimental animals, Δ9-THC or WIN55212-2 produces classical pharmacological effects (analgesia, catalepsy, hypothermia, hypolocomotion), biphasic changes in affect (reward vs. aversion, anxiety vs. anxiety relief), and cognitive deficits (spatial learning and memory, short-term memory). Accumulating evidence indicates that activation of CB1Rs underlies the majority of Δ9-THC or WIN55121-2's pharmacological and behavioral effects. Unexpectedly, glutamatergic CB1Rs preferentially underlie cannabis action relative to GABAergic CB1Rs. Functional roles for CB1Rs expressed on astrocytes and mitochondria have also been uncovered. In addition, Δ9-THC or WIN55212-2 is an agonist at CB2R, GPR55 and PPARγ receptors and recent studies implicate these receptors in a number of their CNS effects. Other receptors (such as serotonin, opioid, and adenosine receptors) also modulate Δ9-THC's actions and their contributions are detailed. This chapter describes the neural mechanisms underlying cannabis action, which may lead to new discoveries in cannabis-based medication development for the treatment of cannabis use disorder and other human diseases.


Enumeration of CD4+ T-cells using a portable microchip count platform in Tanzanian HIV-infected patients.
CD4(+) T-lymphocyte count (CD4 count) is a standard method used to monitor HIV-infected patients during anti-retroviral therapy (ART). The World Health Organization (WHO) has pointed out or recommended that a handheld, point-of-care, reliable, and affordable CD4 count platform is urgently needed in resource-scarce settings. HIV-infected patient blood samples were tested at the point-of-care using a portable and label-free microchip CD4 count platform that we have developed. A total of 130 HIV-infected patient samples were collected that included 16 de-identified left over blood samples from Brigham and Women's Hospital (BWH), and 114 left over samples from Muhimbili University of Health and Allied Sciences (MUHAS) enrolled in the HIV and AIDS care and treatment centers in the City of Dar es Salaam, Tanzania. The two data groups from BWH and MUHAS were analyzed and compared to the commonly accepted CD4 count reference method (FACSCalibur system). The portable, battery operated and microscope-free microchip platform developed in our laboratory (BWH) showed significant correlation in CD4 counts compared with FACSCalibur system both at BWH (r = 0.94, p<0.01) and MUHAS (r = 0.49, p<0.01), which was supported by the Bland-Altman methods comparison analysis. The device rapidly produced CD4 count within 10 minutes using an in-house developed automated cell counting program. We obtained CD4 counts of HIV-infected patients using a portable platform which is an inexpensive (<$1 material cost) and disposable microchip that uses whole blood sample (<10 µl) without any pre-processing. The system operates without the need for antibody-based fluorescent labeling and expensive fluorescent illumination and microscope setup. This portable CD4 count platform displays agreement with the FACSCalibur results and has the potential to expand access to HIV and AIDS monitoring using fingerprick volume of whole blood and helping people who suffer from HIV and AIDS in resource-limited settings.


Practical considerations for quantitative light sheet fluorescence microscopy.
Fluorescence microscopy has evolved from a purely observational tool to a platform for quantitative, hypothesis-driven research. As such, the demand for faster and less phototoxic imaging modalities has spurred a rapid growth in light sheet fluorescence microscopy (LSFM). By restricting the excitation to a thin plane, LSFM reduces the overall light dose to a specimen while simultaneously improving image contrast. However, the defining characteristics of light sheet microscopes subsequently warrant unique considerations in their use for quantitative experiments. In this Perspective, we outline many of the pitfalls in LSFM that can compromise analysis and confound interpretation. Moreover, we offer guidance in addressing these caveats when possible. In doing so, we hope to provide a useful resource for life scientists seeking to adopt LSFM to quantitatively address complex biological hypotheses.


Light-Dark Open Field (LDOF): A novel task for sensitive assessment of anxiety.
Pre-clinical studies of psychiatric disorders often include a measure of anxiety-like behavior. Several tasks exist that serve this purpose, but because anxiety is complex with a myriad of anxiogenic stimuli, researchers are often compelled to use multiple tasks. The Light-Dark Open Field (LDOF) combines concepts from two such tasks, Light-Dark Box and Open Field, into one task with the synergistic effect of enhanced discrimination of anxiety-like behavior. Our goal was to increase the sensitivity of the Open Field task with the addition of a shadow, conceptually similar to the Light-Dark Box, to detect concealed differences even under bright light, which is highly anxiogenic. The resulting LDOF allows assessment of anxiety due to bright light and open space simultaneously, while retaining the ability to assess the impact of each with custom indices. In addition, it maintains all the advantages and measures of the Open Field. Using custom created indices from measures collected in the LDOF one can assess anxiety induced by light, open space, or light and open space combined and thus elucidate anxiety-inducing factors. Using two strains of rats: an outbred strain, Sprague-Dawley (SD), and a strain that exhibits high trait anxiety, Lewis rats, we found that increased discrimination for anxiety-like behavior can be achieved with the Light-Dark Open Field. The LDOF allows researchers to extract the traditional measures of an Open Field, including valuable information about locomotion and habituation while adding a further layer of discrimination with the light-dark component. Because the LDOF is a combination of two different tests, it saves time compared to running multiple experiments in series that then need to be counterbalanced to reduce artefacts and task order effects. In addition, it detects differences even when traditional tasks of anxiety have reached their ceiling sensitivity (i.e. EPM under bright light conditions). We present the Light-Dark Open Field: a simple modification of an existing Open Field apparatus that incorporates aspects of the Light-Dark Box with the addition of a shadow. The shadow (Dark Perimeter) allows for increased discrimination in detecting anxiety-like behaviors. Comparison of anxiety-like behavior between Lewis and SD rats allowed us to develop the construct and face validity of the LDOF as well as demonstrate its sensitivity even under bright light conditions. In addition, we developed 3 indices that allow one to parse out, from one set of data, the effect of two anxiogenic stimuli- bright light and open space.


A review of the evidence for the use of metformin in the treatment of metabolic syndrome caused by antipsychotics.
Psychiatric patients requiring therapy with antipsychotics have a greater incidence of becoming overweight or obese compared with the general population. Many of these patients are often treated with second-generation (atypical) antipsychotics (SGAs), which are associated with weight gain, dyslipidaemia, and other metabolic derangements. The most important and first line of treatment for the metabolic syndrome is lifestyle changes including diet and exercise. However, other approaches like the use of medication (e.g. Metformin) have been also used, mainly when the lifestyle changes are difficult to achieve. Therefore, the treatment of antipsychotic-induced weight gain with metformin may be an option after the lifestyle and dietary changes fail. The use of metformin is still experimental and off license regarding the treatment of metabolic syndrome in Psychiatric patients, however we wished to assess the evidence for its use. Our study is a literature based research. For our research we reviewed 12 Pubmed published articles from 2006 to 2013. Metformin have been reported to counteract effectively antipsychotic-induced body weight gain and has been demonstrated to improve glycaemic control and promote a moderate weight loss in both diabetic and non-diabetic subjects. Metformin use appears to be a benefit when started early in the course of treatment and mostly in young adults newly exposed to antipsychotic drugs.


Helicobacter pylori-Induced DNA Damage Is a Potential Driver for Human Gastric Cancer AGS Cells.
Helicobacter pylori is a major cause of gastric cancer. This study was aimed to explore the characteristic of DNA damage induced by H. pylori infection in gastric cancer AGS cells. After infection with H. pylori, the reactive oxygen species (ROS) levels in AGS cells were significantly higher than those in the uninfected cells. Cells with longer comet tails were detected after infection with H. pylori. The number of apurinic/apyrimidinic endonuclease 1- and phosphorylated H2AX-positive cells was significantly increased compared with the number of negative control cells. The expression of pChk1 and pChk2 was significantly upregulated by H. pylori infection. Cell growth was inhibited after H. pylori infection. All these results were dose dependent. The cell alterations were more significant upon infection with H. pylori at a multiplicity of infection (MOI) of 100:1 than at an MOI of 50:1. H. pylori infection can induce DNA single-strand breaks, DNA double-strand breaks, and cell cycle checkpoint activation after ROS generation in the gastric cancer cell line AGS, which is a potential driver for gastric cancer.